【公開日：2023.07.31】【最終更新日：2023.05.16】
課題データ / Project Data
課題番号 / Project Issue Number
22UT0028
利用課題名 / Title
シャペロニンタンパク質GroELの自己組織化
利用した実施機関 / Support Institute
東京大学
機関外・機関内の利用 / External or Internal Use
内部利用（ARIM事業参画者以外）/Internal Use (by non ARIM members)
技術領域 / Technology Area
【横断技術領域 / Cross-Technology Area】（主 / Main）計測・分析/Advanced Characterization（副 / Sub）-
【重要技術領域 / Important Technology Area】（主 / Main）次世代バイオマテリアル/Next-generation biomaterials（副 / Sub）-
キーワード / Keywords
Proteins, GroEL, Self-Assembly, Nanomaterials, Transmission Electron Microscopy,電子顕微鏡/Electron microscopy,バイオアダプティブ材料/ Bioadaptive materials,DDSマテリアル/ DDS material,生体イメージング/ In vivo imaging,細胞・組織再生誘導材料/ Materials for inducing cell and tissue regeneration
利用者と利用形態 / User and Support Type
利用者名（課題申請者）/ User Name (Project Applicant)
藤原 潤紀
所属名 / Affiliation
東京大学
共同利用者氏名 / Names of Collaborators in Other Institutes Than Hub and Spoke Institutes
ARIM実施機関支援担当者 / Names of Collaborators in The Hub and Spoke Institutes
木村鮎美,寺西亮佑
利用形態 / Support Type
（主 / Main）技術補助/Technical Assistance（副 / Sub）,機器利用/Equipment Utilization
利用した主な設備 / Equipment Used in This Project
UT-010：クライオ透過型/透過走査型電子顕微鏡
UT-011：有機材料ハイコントラスト透過型電子顕微鏡
報告書データ / Report
概要（目的・用途・実施内容）/ Abstract (Aim, Use Applications and Contents)
Supramolecular nanostructures, including nanoparticles, vesicles, and 3D networks, formed through the self-assembly of the chaperonin protein GroEL were visualized by transmission electron microscopy (TEM), including under cryogenic conditions (cryo-TEM).
実験 / Experimental
The chaperonin protein GroEL, a model biomolecular machine, was site-specifically functionalized with DNA and a range of other polymers and small molecules through genetic and chemical modifications. These functionalized moieties were incorporated to program the self-assembly of the GroEL species into the various supramolecular nanostructures. The obtained nanostructures were observed by TEM in the dry state (JEM-1400) after uranyl acetate staining to see fine structural details, down to the orientation of individual GroEL units, and also in the cryogenic state (JEM-2100F) in order to preserve the structure as it exists in the dispersed state.
結果と考察 / Results and Discussion
The structures of gold nanoparticles (NPs) with dense shells of GroEL on their surface were observed in the dry state, and the visualization of the characteristic stripe pattern of GroEL confirmed their homeotropic alignment on the NP surface. Separately, GroEL vesicles were formed through the self-assembly of GroEL species functionalized anisotropically with polyethylene glycol (PEG), a polymer showing salt- and temperature-responsive hydrophilic-to-hydrophobic transitions. The assembled nanostructures were visualized by cryo-TEM, with and without ammonium molybdate staining, supporting the determination of a hollow monolayer structure. The structures were also visualized by dry-state TEM, showing individual GroEL units.
図・表・数式 / Figures, Tables and Equations
その他・特記事項（参考文献・謝辞等） / Remarks(References and Acknowledgements)
成果発表・成果利用 / Publication and Patents
論文・プロシーディング（DOIのあるもの） / DOI (Publication and Proceedings)
口頭発表、ポスター発表および、その他の論文 / Oral Presentations etc.
- ○Kiyoshi Morishita, PK Hashim, Haruaki Yanagisawa, Masahide Kikkawa, Hideki Taguchi, Takuzo Aida, Biomolecular Machines as Functional Building Blocks in Programmed Nanostructures. Chemical Society of Japan 103rd Annual Meeting, Noda, Japan, March 23, 2023.
特許 / Patents
特許出願件数 / Number of Patent Applications：0件
特許登録件数 / Number of Registered Patents：0件